CD370 (Clec9A) Mouse anti-Human, PerCP-eFluor 710, Clone: 9A11, eBioscience
Mouse Monoclonal Antibody
Manufacturer: Invitrogen 46370941
This monoclonal antibody 9A11 reacts with human CD370, also known as C-type lectin family member 9A (Clec9A) and DNGR1. This type II transmembrane glycoprotein is present at the cell surface in the form of a homodimer. CD370 is a cellular receptor expressed by myeloid cells that recognizes F-actin, a dead cell-associated antigen that is exposed in necrotic cells. Upon ligation, CD370 recruits Syk kinase to the ITAM-like motif in its intracellular tail and facilitates endocytosis of antigenic material that is subsequently processed and cross-presented to CD8+ T cells. Human CD370 is expressed by CD141+ (BDCA3+) dendritic cells and, to a lesser degree, by CD14+ CD16- monocytes. Unlike in mice, human plasmacytoid dendritic cells do not express CD370. This 9A11 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Filtration: 0.2 µm post-manufacturing filtered. CLEC9A is a group V CTLR (C-type lectin-like receptor) that functions as an activation receptor and is expressed on myeloid lineage cells. This protein is also expressed at the cell surface and acts as glycosylated dimers which mediate endocytosis, but not phagocytosis (Poulin LF et al, 2010). It facilitates antigen uptake and presentation, and may provide a suitable target for antibody-mediated antigen delivery. CLEC9A is important for conveying information from necrotic cells to T-cells and defines a pathway by which adaptive immune responses can be initiated in the absence of infection. It recruits Syk to mediate sensing of necrosis by the principal DC subset involved in regulating cross priming to cell-associated antigens. Signaling via this kinase occurs through a novel pathway involving CARD9 which induce a variety of cellular responses including the induction of cytokines (such as TNF, IL-6, IL-10, IL-23, and IL-2), the respiratory burst, and the production of Arachidonic acid. In addition, it also induces TH17 (T-helper type 17) adaptive responses in vivo. Expression of CLEC9A is highly restricted in peripheral blood, being detected only on BDCA3+ DCs and on a small subset of CD14+ CD16- monocytes. CLEC9A serves as a new marker to distinguish subtypes of DCs.
|PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide|
|4° C, store in dark, DO NOT FREEZE!|
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